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Chapter 4 Influence of Environmental Factors Environmental Factors 1. Nutrients 2. Temperature 3. pH-Buffer capacity 4. Mixing 5. Presence of alternative electron acceptors (SO42-, NO3-, etc.) 1 Environmental factors affect ... Specific growth rate Decay rate Gas production rate Substrate utilization rate Start-up Response to change in input 1. Nutrients Nutrients supply the basic cellular building blocks for growth and ensure the cell is able to synthesize the enzymes and cofactors that drive the biochemical and metabolic reactions. According to the relative quantities required by the cell, nutrients can be divided into two groups; Macro-nutrients Micro-nutrients 2 Macronutrients Both macro- and micronutrients have to be present in an available form in the growth environment to allow effective uptake. Ideally, nutrient levels should be in excess of the optimum concentrations required. Because anaerobic bacteria can be severely inhibited by even slight nutrient deficiencies. However, many essential nutrients can become toxic when present in high concentrations. Macronutrients: Nitrogen A rough estimate of the theoretical amount of macronutrients (N, P and S) can be derived from elemental composition of bacterial cells within anaerobic sludge. Emprical formula of biomass: C5H7O2N then; 3 - 6 kg N /1000 kg of COD consumed or 0.5 -10 kg N /60 m3 of CH4 produced COD:N ratio ~ 350:7 (High organic loading) COD:N ratio ~ 1000:7 (Low organic loading) COD:N ratio ~ 200:5 (Recommended) 3 Macronutrients: Nitrogen Most common nitrogen forms; ammonia (NH3), nitrate (NO3-), nitrite (NO2-), nitrogen gas (N2). NH3 is the most readily utilized inorganic forms of nitrogen, existing in the reduced state that is required for anabolic metabolism and an uncharged state that facilitates cellular uptake. Macronutrients: Phosphorus N:P ratio ~ 7:1 (Recommended) Then COD/N/P ratio ~ 300:7:1 The usual forms of āPā in aqueous solution include orthophosphate, polyphosphate & organic phosphate. The orthophosphates are immediately available for biological metabolism without further modification. Organic phosphates must generally be hydrolysed by the cell to release inorganic phosphate before use. 4 Macronutrients: Sulfur In addition to N and P, the sulfur (S) requirement of anaerobic bacteria should also be satisfied and this can be supplied as sulfur, sulfide, sulfite, thiosulfate, sulfate or amino acids (cysteine and methionine). Optimum anaerobic digester concentrations of S have been reported between 0.001 and 1.0 mg/l. Composition of methanogens Typical elemental composition (mg/l) of methanogens 5 Micronutrients Nutrient Ca Concentration required (mg/l) Effects on digestion 100-200 Granulation and increase in activity Mg 75-150 Granulation and increase in activity Na 100-200 Increase in activity Fe 20-100 Increase in activity and precipitation of sulphide K 200-400 Increase in activity Ba 0.01-0.1 Co 20 Vitamin B12 dependent W - Formate dehydrogenase Se 0.8 SO42- 0.1-10 Divalent cation effect hence good granulation Formate dehydrogenase, glycine reductase, hydroxylase, and dehydrogenase dependent Sulfur source of cell synthesis 2. Temperature The most influential environmental factors as it controls the activity of all microorganisms. A rise in temperature leads to an increase in the rate of biochemical and enzymatic reactions within cells, causing increased growth rates. Above a specific temperature, which is characteristic of each species, this phenomenon gives way to inhibition and then mortality, as the proteins and structural components of the cell become irreversibly denatured. 6 Temperature and Growth The effect of temperature on growth rate Temperature ranges for growth 7 Temperature ranges for AD Psychrophilic AD is favoured by organisms having a temperature optimum at 15-20oC. Although it is not as efficient as mesophilic and thermophilic AD, it has desirable economic trade-offs in temperate climates. Mesophilic AD with a temperature optimum at 30-37oC is the most commonly employed in engineered processes of anaerobic treatment. Thermophilic AD with an optimum at 55-60oC is more efficient than mesophilic AD. Mesophilic vs Thermophilic Thermophilic reactors can accept higher organic loading rates and produce lower quantities of sludge. Mesophilic reactors are often more stable. Thermophilic reactors require more energy to heat the reactor Thermophilic reactors produce high concentrations of VFA in their effluent. Thermophilic AD is an attractive option for treating warm industrial effluents and slurries of relatively constant composition. 8 Anaerobic Sludge Digestion Temperature (oC) 60 50 Thermophilic 40 Mesophilic 30 20 Psychrophilic 10 20 40 60 80 100 HRT, days Influence of temperature Compared to aerobic processes which are relatively robust to temp. variations, AD is sensitive to sudden temperature fluctuations Temp. changes as small as 1-2oC have significant adverse effects on process performance particularly when changes occur rapidly (<2 hrs). If bacteria become adversely affected by temperature variations, several days or even weeks may be required to restore a healthy population once again. 9 3. pH-Buffer capacity For an efficient methanogenic digestion a suitable and stable pH has to be maintained within the digester. pH has critical influences on; Microorganisms (esp. Methanogens) responsible for AD Biochemistry of AD process Alkalinity buffering and Chemical reactions affecting the solubility and availability of dissolved ions. Effect of pH Best pH range appears to be around neutrality, while 6.5-8.0 is generally believed to be optimal. The effect of pH on biogas production. 10 Reactions affecting pH Four major chemical and biochemical reactions that influence the pH of an AD are: Ammonia (NH3) consumption and release Volatile fatty acid production and consumption Sulfide (S2-) release by dissimilatory reduction of sulfate (SO42-) or sulfite (SO32-) Conversion of neutral carbonaceous organic carbon to methane (CH4) and carbon dioxide (CO2) Controlling the pH In AD, pH reduction can be countered by formation of bicarbonate alkalinity and consumption of VFAs by methanogens. Consumption of VFAs is dependent on the equilibrium between rapid growing acidogens and slow growing methanogens. This equilibrium can be easily upset by changes in the operational or environmental conditions. 11 Controlling the pH If the balance is disturbed and therefore pH starts to drop because of VFAs accumulation. Stop feeding the reactor to give the methanogens sufficient time to consume excess VFAs and raise the pH value to an acceptable level. OR Dose the reactor with alkali, i.e. NaOH, Na2CO3 or NaHCO3 in order to raise the pH or provide additional buffering capacity. In some cases both options are used simultaneously. Controlling the pH The amount of alkalinity required to accommodate VFA increases in AD depends on many factors. Well- established anaerobic reactors treating typical organic loads are likely to contain alkalinity in the range 2000 to 3000 mg/l as CaCO3. This level of alkalinity will impart an improved resistance to acidification caused by short-term fluctuations in feed composition. 12 4. Mixing AD comprises an inherent degree of mixing from the continuous rise of biogas bubbles within the reactor, This mixing is not sufficient for efficient mass transfer. Mixing enhances the AD process by distributing microorganisms, substrate, and nutrients throughout the digester as well as equalizing temperature. Mixing also provides for rapid hydrolysis of wastes by allowing the hydrolytic bacteria to attack a much larger surface area. Types of Mixing The level and type of mixing also affects; Growth rate and distribution of bacteria within the sludge Substrate availability and utilization rates Granule formation and Biogas production. Mixing can be enhanced using; Mechanical devices (paddles, turbines and propellers) Hydraulic shear force (feed recycle) Biogas recirculation 13 Advantages of Mixing Eliminating or reducing scum build-up Eliminating thermal stratification or localized pockets of depressed temperature Maintaining digester sludge chemical and physical uniformity throughout the tank Rapid dispersion of metabolic wastes (products) produced during substrate digestion Rapid dispersion of any toxic materials entering the tank (minimizing toxicity) Prevent deposition of grit Excessive Mixing Excessive mixing may lead to a reduction in reactor performance by disrupting the flocs and granules containing active anaerobic microorganisms. Result in short-circuiting of the reactor, leading to unconverted substrate appearing in the reactor effluent. Lead to the formation of smaller flocs, which have poor settling characteristics. The kind of mixing equipment and amount of mixing varies with the type of reactor and the solids content in the digester. 14 5. Presence of alternative electron acceptors Under anaerobic conditions sulfate (SO42-) and sulfite (SO32-) is reduced to sulfide (S2-) by SRB. The SRB utilize electron-donating substrates present in wastewater for the reduction of sulfate. The substrates are either partially oxidized to acetate or fully oxidized to CO2. Sulfate behaves as an alternative electron acceptor to support anaerobic respiration. Sulfate redution lower the CH4 yield per kg organic waste Biogas treatment is required to remove corrosive H2S 5. Presence of alternative electron acceptors Denitrification is an anoxic process in which either an organic or inorganic electron-donating substrates are oxidized at the expense of reducing nitrate (NO3-) or nitrite (NO2-) to dinitrogen gas (N2). Denitrifiers have the ability to utilize a variety of fermentative/methanogenic substrates therefore these microorganisms compete for the same substrate(s) such as glucose, VFAs and H2. Propionate is the most preferred VFAs as carbon source by denitrifiers. 15